Pharmaceutical composition for treating female infertility and preparation method thereof

Abstract

The invention provides a pharmaceutical composition for treating female infertility and a preparation method thereof, and belongs to the technical field of traditional Chinese medicines. Aiming at the treatment of female infertility caused by ovulation dysfunction, the pharmaceutical composition for treating female infertility is obtained by combining nine medicines, namely gallnut, goosegrass herb, rosa cymosa, caragana, euonymus alatus, cortex lycii radicis, melastoma dodecandrum root, fructus rosae laevigatae and radix fici simplicissimae. According to the pharmaceutical composition disclosed by the invention, all the medicines are synergistically matched, so that good effects of promoting follicular development, improving follicular atresia and apoptosis and recovering sex hormone level are achieved, and the pharmaceutical composition has a good effect in the aspects of relieving female infertility and improving female reproductive disorder.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a pharmaceutical composition for treating female infertility and a preparation method thereof. Background Art

[0002] Infertility refers to the failure of a couple of childbearing age to conceive after living together for more than one year (some standards are two years), having normal sexual life, and not taking any contraceptive measures. The incidence of infertility is very high in modern society, which has a great impact on the patients themselves and their families.

[0003] Infertility is divided into infertility and infertility syndrome. The main causes of female infertility include: fallopian tube obstruction, hydrops; uterine lesions, such as uterine fibroids, adenomyosis, intrauterine adhesions, etc.; ovulation dysfunction, such as polycystic ovary syndrome, premature ovarian failure, hyperprolactinemia, etc. In addition, systemic diseases such as chromosomal abnormalities, reproductive organ malformations, and lupus erythematosus caused by genes can also affect normal reproductive function.

[0004] Western medicine treatment of infertility is mainly divided into drug therapy and surgical treatment. Commonly used drugs include prednisone acetate, L-carnitine, and multivitamins. Prednisone acetate is a glucocorticoid drug with a wide range of applications. It has anti-inflammatory, anti-allergic, and anti-shock effects, and has a good therapeutic effect on infertility caused by positive antisperm antibodies. However, as a hormone drug, it may produce certain adverse reactions, such as nausea and vomiting, rash, diarrhea, weight gain, etc.; it may also stimulate the nervous system, causing adverse reactions such as overexcitement, agitation, depression, and depression. In addition, long-term use of Western medicine will increase the metabolic burden of the liver, and in severe cases, it can also cause liver damage, causing more serious harm to the patient's body.

[0005] There are several main types of TCM syndrome differentiation for infertility: ① Kidney Yang Deficiency Type, the patient's Chong and Ren meridians are out of balance, the blood sea is not solid, resulting in insufficient kidney yang, and the life gate fire is weak. Men often experience symptoms such as soreness of the waist and knees, impotence, premature ejaculation, frequent urination, etc., while women have symptoms such as scanty menstruation, dull color, and low libido. ② Kidney Yin Deficiency Type, the patient's yin blood is insufficient, and the body fluid is severely depleted, unable to nourish the whole body. Symptoms such as restlessness, headache, dizziness, general fatigue, night sweats, and tinnitus may occur. ③ Liver Depression Type, the patient's emotions are uncomfortable and his mood is in a low state for a long time, resulting in liver depression and qi stagnation, poor blood and qi circulation, and systemic yin and yang imbalance, which will cause irritability, decreased libido, depression, lower abdominal discomfort, insomnia and dreaminess. ④ Phlegm-dampness type, more common in obese patients, phlegm blocks the qi, meridians are blocked, and sperm cannot be collected to conceive. Women's blood and qi are out of balance, resulting in infrequent menstruation or even amenorrhea, with specific manifestations of coughing with a lot of sputum, white and sticky sputum, dry mouth with little fluid, thick and greasy tongue coating, etc. ⑤ Qi stagnation and blood stasis type. The patient's qi and blood do not flow smoothly, blood stasis blocks the uterine vessels, women have less menstrual blood, and men's essence is blocked, resulting in infertility.

[0006] According to TCM syndrome differentiation and treatment, the kidney is the origin of the human body, and the occurrence of many diseases in the body is closely related to the obstruction or decline of kidney function. The kidney has the functions of storing essence, controlling water, and absorbing qi. Most of the essence of the human body is stored in the kidney, which can prevent the loss of essence for no reason. The kidney can also regulate the metabolism of human water, participate in the blood circulation of the whole body, and promote the harmony of yin and yang. The occurrence of infertility is likely to be related to renal insufficiency. The deficiency of kidney qi leads to the accelerated loss of essence and qi, and the obstruction of blood and qi circulation throughout the body, which makes the imbalance of yin and yang in the body affect the reproductive function. The methods of TCM treatment of infertility mainly include TCM conditioning, injection therapy, etc.

[0007] She doctors believe that human life is dominated by the "spirits" of the six organs, namely the heart, liver, lungs, spleen, kidneys and gallbladder (called the "six spirits"). If the body's qi and blood are vigorous and the tendons and veins are unobstructed, life activities are normal; if the qi and blood are unbalanced or exhausted, the tendons and veins are blocked, and the plague evil erodes, diseases will occur. Female infertility is also caused by qi and blood imbalance, blood stagnation, and lack of nourishment to the cells and veins. Its treatment principle is mainly to regulate the balance of the human body.

[0008] At present, there are many prescriptions for treating female infertility using traditional Chinese medicine on the market, such as Zishen Yutai Pills and Yougui Pills for patients with spleen and kidney deficiency, Kuntai Capsules for patients with heart-kidney disharmony, Xiaoyao Pills for patients with liver qi stagnation, Dingkun Dan for patients with qi and blood deficiency, and Shaofu Zhuyu Pills for patients with uterine stasis. However, the therapeutic effects of these traditional Chinese medicines are not ideal, and most of the prescriptions use a large amount of kidney and liver tonics, which are not easy to absorb and are easy to get angry after eating, which also greatly reduces the efficacy. Therefore, there is a need for a traditional Chinese medicine with better efficacy for treating female infertility. Summary of the invention

[0009] The purpose of the present invention is to provide a pharmaceutical composition for treating female infertility and a preparation method thereof. The various herbs in the pharmaceutical composition of the present invention cooperate synergistically to promote follicular development, improve follicular atresia and apoptosis, and restore sex hormone levels, and have good efficacy in alleviating female infertility and improving female reproductive disorders.

[0010] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0011] The invention provides a pharmaceutical composition for treating female infertility. The pharmaceutical composition is prepared from the following raw materials in parts by weight: 30-60 parts of gallnut, 20-40 parts of goosegrass, 25-50 parts of small-fruited rose, 20-40 parts of caragana, 20-40 parts of euonymus laxative, 25-50 parts of lycium bark, 20-40 parts of euonymus sparsely flowered, 20-40 parts of euonymus laxative, 20-40 parts of euonymus root, 20-40 parts of Rosa laevigata, and 30-60 parts of pheasant.

[0012] Preferably, the pharmaceutical composition is prepared from the following raw materials in parts by mass: 30 parts of Galla chinensis, 20 parts of Eleocharis chinensis, 25 parts of Rosa microcarpa, 20 parts of Caragana, 20 parts of Euonymus laxative, 25 parts of Lycium bark, 20 parts of Radix Euonymus japonicus, 20 parts of Rosa laevigata, and 30 parts of Prunus mume.

[0013] The present invention also provides a method for preparing a pharmaceutical composition for treating female infertility, comprising the following steps:

[0014] After all the drugs are mixed, they are soaked in water, decocted for the first time and filtered, the drug residues are decocted for the second time and filtered, and the filtrates are combined, concentrated, dried, and sterilized to obtain a drug composition for treating female infertility.

[0015] Preferably, the mass ratio of the traditional Chinese medicine to water during the soaking in water is 1:5-8, and the soaking time in water is 10-12 hours.

[0016] Preferably, the first decoction time is 1 to 3 hours; the mass ratio of traditional Chinese medicine to water during the second decoction is 1:5 to 8, and the decoction time is 1 to 3 hours.

[0017] Preferably, the pharmaceutical composition for treating female infertility can be prepared into decoction, capsule, paste, powder or pill.

[0018] The beneficial effects of the present invention compared with the prior art are:

[0019] The present invention is aimed at treating female infertility caused by ovulation dysfunction, and nine medicines including gallnut, smilax china, small-fruited rose, caragana, euonymus laxative, lycium bark, radix rutaecarpa, rosa laevigata, and pheasant are selected for compatibility to obtain a pharmaceutical composition for treating female infertility. The various medicines in the pharmaceutical composition of the present invention cooperate synergistically, play a good role in promoting follicular development, improving follicular atresia and apoptosis, and restoring sex hormone levels, and have good efficacy in alleviating female infertility and improving female reproductive disorders. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0021] Figure 1 This is a flow chart for detecting the effect of the pharmaceutical composition for treating female infertility in Test Example 1 of the present invention;

[0022] Figure 2The weight changes of mice in the POF and PCOS groups before and after modeling and drug administration in Experimental Example 1 of the present invention;

[0023] Figure 3 The follicle development of mice in the POF group in Experimental Example 1 of the present invention;

[0024] Figure 4 The follicle development of mice in the PCOS group in Experimental Example 1 of the present invention;

[0025] Figure 5 The HE staining results of the uterus of mice in the POF and PCOS groups in Experimental Example 1 of the present invention;

[0026] Figure 6 This is the sex hormone secretion of mice in the POF and PCOS groups in Experimental Example 1 of the present invention. DETAILED DESCRIPTION

[0027] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0028] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0029] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0030] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0031] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0032] The present invention prepares a pharmaceutical composition for treating female infertility, and the specific preparation method is as follows:

[0033] Weigh the medicines according to the weight proportions of 30-60 parts of Galla chinensis, 20-40 parts of Eleocharis truncatula, 25-50 parts of Rosa microcarpa, 20-40 parts of Caragana, 20-40 parts of Euonymus laxative, 25-50 parts of Lycium bark, 20-40 parts of Radix Euonymus, 20-40 parts of Radix Euryale, 20-40 parts of Rosa laevigata, and 30-60 parts of Phragmites australis and mix them evenly.

[0034] Weigh the mixed medicine, soak it in 5-8 times the amount of water for 10-12 hours, then decoct it for the first time for 1-3 hours, filter it, and collect the first extract. Decoction the residue again with 5-8 times the amount of water for 1-3 hours, filter it, collect the second extract, combine the filtrate, concentrate, dry, and sterilize it to obtain a pharmaceutical composition for treating female infertility.

[0035] Example 1

[0036] Embodiment 1 of the present invention provides a method for preparing a pharmaceutical composition for treating female infertility, which specifically comprises the following steps:

[0037] Weigh the medicines according to the weight proportions of 30 parts of Galla chinensis, 20 parts of Eleocharis truncatula, 25 parts of Rosa microcarpa, 20 parts of Caragana, 20 parts of Euonymus laxative, 25 parts of Cortex Lycii, 20 parts of Radix Euonymus, 20 parts of Radix Dioscoreae, 20 parts of Rosa laevigata, and 30 parts of Psoralea corylifolia, and mix them evenly.

[0038] Weigh 500.16g of the mixed medicine, soak it in 6 times the amount of water as the medicinal material for 10 hours, then transfer all of it to a 5L flat-bottom flask, reflux extract for 2 hours by decoction method, filter, and collect the first extract. The medicinal residue is reflux extracted again with 6 times the amount of water for 2 hours, and the second extract is collected by filtration. Combine the two extracts and distill under reduced pressure (68°C) in a pre-weighed rotary evaporator to obtain a thick paste, and freeze-dry the thick paste obtained after reduced pressure concentration to obtain a pharmaceutical composition for treating female infertility.

[0039] Example 2

[0040] Embodiment 2 of the present invention provides a method for preparing a pharmaceutical composition for treating female infertility, which specifically comprises the following steps:

[0041] Weigh the medicines according to the weight proportions of 60 parts of Galla chinensis, 35 parts of Eleocharis truncatula, 25 parts of Rosa microcarpa, 35 parts of Caragana, 35 parts of Euonymus laxative, 25 parts of Cortex Lycii, 35 parts of Radix Euryale ferox, 35 parts of Rosa laevigata, and 60 parts of Psoralea corylifolia, and mix them evenly.

[0042] Weigh 500.16g of the mixed medicine, soak it in 8 times the weight of the medicinal material in water for 10 hours, then transfer all of it to a 5L flat-bottom flask, reflux extract for 3 hours by decoction, filter, and collect the first extract. The medicinal residue is reflux extracted again with 6 times the weight of water for 2 hours, and the second extract is collected by filtration. Combine the two extracts and distill under reduced pressure (68°C) in a pre-weighed rotary evaporator to obtain a thick paste, and freeze-dry the thick paste obtained after reduced pressure concentration to obtain a pharmaceutical composition for treating female infertility.

[0043] Example 3

[0044] Example 3 of the present invention uses the pharmaceutical composition for treating female infertility prepared in Example 1 to prepare a pharmaceutical composition capsule for treating female infertility, and the specific steps are as follows:

[0045] The thick paste in Example 1 was placed in a drying oven and dried at 130° C., crushed into 300-400 mesh fine powder, and sterilized to prepare capsules.

[0046] Test Example 1

[0047] Test Example 1 of the present invention Figure 1 The process shown detects the therapeutic effect of the pharmaceutical composition for treating female infertility prepared in Example 1, which specifically includes the following

[0048] (1) Establishment of POF mouse model:

[0049] Sixteen 8-week-old C57BL / 6 female mice were randomly divided into 4 groups, with 4 mice in each group, namely negative control group, positive control group, low-dose group of the pharmaceutical composition of Example 1, and high-dose group of the pharmaceutical composition of Example 1. The model mice were all intraperitoneally injected with cyclophosphamide (75 mg / Kg) once, and the negative control group was injected with an equal volume of phosphate buffer once. All groups were fed with free food (high-pressure maintenance feed) and drinking water for 7 days, with a temperature of 25±3°C and a relative humidity of 50-60%. After 7 days, the pharmaceutical composition of Example 1 was gavaged every day (twice a day, with an interval of more than 4 hours), 225 mg / mL in the low-dose group and 450 mg / mL in the high-dose group, and the drug was administered continuously for 7 days at 0.2 ml / 10g according to body weight. The negative and positive control groups were gavaged with an equal amount of double distilled water. During the period, the initial weight of the mice, the weight after modeling, and the weight after seven days of administration were recorded, and One-way ANOVA statistical analysis was performed. The results are as follows Figure 2 shown.

[0050] (2) Establishment of PCOS mouse model:

[0051] Sixteen 21-day-old C57BL / 6 female mice were randomly divided into 4 groups, namely a negative control group, a positive control group, a low-dose group of the pharmaceutical composition of Example 1, and a high-dose group of the pharmaceutical composition of Example 1. The model mice were subcutaneously injected with DHEA (6 mg / 100 g, dissolved in sesame oil) for 20 consecutive days, and the negative control group was subcutaneously injected with an equal volume of sesame oil every day. The feeding conditions of each group were the same as those of the POF mice. After 20 days, they were gavaged with the pharmaceutical composition of Example 1 every day (twice a day, with an interval of more than 4 hours), 225 mg / mL in the low-dose group and 450 mg / mL in the high-dose group. The drug was administered at 0.2 ml / 10 g according to body weight for 7 consecutive days, and subcutaneous injections were continued during this period. The negative and positive control groups were gavaged with an equal amount of double distilled water. During this period, the initial weight of the mice, the weight after modeling, and the weight after seven days of administration were recorded, and One-way ANOVA statistical analysis was performed. The results are as follows Figure 2 shown.

[0052] One-way ANOVA statistical analysis: "ns" indicates that there is no significant difference between the other groups and the negative control group, "*(P<0.05)"

[0053] Figure 2 The results showed that the physical development of mice in each group was comparable before and after modeling and prescription treatment, except that the body weight of mice in the POF positive control group was lower, indicating that the pharmaceutical composition of the present invention can improve the body weight maldevelopment caused by POF modeling.

[0054] (3) Follicular development of mice in the POF and PCOS groups

[0055] The ovaries of mice in each group were fixed with 10% formaldehyde solution at room temperature for 12 h, dehydrated with gradient ethanol and embedded in paraffin. The 4 μm thick tissue sections were stained with HE. The results are as follows: Figure 3 , 4 shown.

[0056] Figure 3 The results showed that the ovarian tissue structure of the negative control group in the POF group was intact, the follicles at all levels developed normally, and the corpus luteum and the granulosa cell layer with normal arrangement and stratification were visible. Although the development of follicles at all levels was visible in the positive control group of the POF model, the ovarian tissue was reduced, the follicles were atretic, and the boundaries of the corpus luteum were unclear. However, no atretic follicles were found in the low-dose and high-dose prescription groups of the POF model, the cortex and medulla structures were clear, and more mature follicles (antral follicles) and corpora lutea were seen, indicating that the pharmaceutical composition of the present invention can improve the follicle atresia and apoptosis of the POF model and promote the development of follicles.

[0057] Figure 4It shows that in the PCOS group, the negative control also shows well-developed follicles at all levels, and the granulosa cells are arranged normally. In the positive control group, a large number of secondary follicles and atretic follicles can be seen, the granulosa cells are arranged irregularly, and the number of corpora lutea is very small. In the ovaries of the low-dose prescription treatment group, the follicles at all levels develop normally, and the number of corpora lutea increases significantly. More mature follicles and corpora lutea can be seen in the high-dose group, and mice are mostly in estrus, indicating that the pharmaceutical composition of the present invention can improve the follicular development of the PCOS model and promote estrus in mice.

[0058] (4) Endometrial morphology of mice in POF and PCOS groups

[0059] Ovarian insufficiency and polycystic ovary morphology can lead to thinning of the endometrium, a decrease in the number of glands, and reduced endometrial receptivity. Figure 5 As shown in the figure, the uterus of mice in the POF and PCOS groups was sectioned with paraffin and stained with HE, and it can be seen that the uterine glands of the POF and PCOS models were significantly reduced, the structure was abnormal and unclear, and the glandular atrophy was obvious in the POF positive control, and the nuclear condensation of the glandular epithelial cells was obvious. In the low-dose and high-dose prescription treatment groups, the number of endometrial glands increased significantly, the glandular cavity expanded and contained some secretions, and the matrix showed loose secretory changes.

[0060] (5) Sex hormone secretion in mice of POF and PCOS groups

[0061] The whole blood was collected from the eyeballs of mice in each group, and the serum was obtained by centrifugation at 1000g for 20 minutes and stored at -80℃ for subsequent Elisa kit detection of serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH). One-way ANOVA statistical analysis: "ns" indicates that there is no significant difference between the other groups and the negative control group, and "*(P<0.05), **(P<0.01), ***(P<0.001), ****(P<0.0001)" indicates the significance of the results of the other groups compared with the negative control group.

[0062] Decreased ovarian function is manifested by increased FSH and LH, while polycystic ovary is mainly manifested by increased LH / FSH, low basal FSH levels, and increased or relatively normal LH levels.

[0063] Figure 6 The results showed that in the POF model, the FSH and LH levels of the positive control were significantly higher than those of the negative control group, while the serum FSH and LH levels of the low-dose and high-dose prescription treatment groups were significantly reduced and tended to return to normal.

[0064] In the PCOS model, the FSH level of the positive control was significantly reduced, the LH level was normal, and the LH / FSH ratio was significantly higher than that of the negative control group. After treatment with the low-dose and high-dose prescriptions, the FSH level was significantly restored, and when the LH level was equivalent, the LH / FSH ratio was significantly reduced. Therefore, the pharmaceutical composition of the present invention has a good hormone regulation effect.

[0065] It can be seen from the above examples that the pharmaceutical composition of the present invention can effectively improve the effects of ovarian function damage and polycystic ovary on the female reproductive system, effectively improve the development of follicles at all levels and promote estrus in mice, improve the endometrial receptivity of mice after modeling, and effectively regulate the imbalance of FSH and LH hormone levels caused by ovarian function damage.

[0066] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A pharmaceutical composition for treating female infertility, characterized in that: The pharmaceutical composition is prepared from the following raw materials in parts by weight: 30-60 parts of Galla chinensis, 20-40 parts of Eleutherodactyla serrata, 25-50 parts of Rosa microcarpa, 20-40 parts of Caragana, 20-40 parts of Euonymus laxative, 25-50 parts of Lycium bark, 20-40 parts of Radix Euonymus japonicus, 20-40 parts of Rosa laevigata, and 30-60 parts of Prunus mume 2. The pharmaceutical composition for treating female infertility according to claim 1, characterized in that: The pharmaceutical composition is prepared from the following raw materials in parts by weight: 30 parts of Galla chinensis, 20 parts of Eleutherodactyla serrata, 25 parts of Rosa microcarpa, 20 parts of Caragana, 20 parts of Euonymus laxative, 25 parts of Lycium bark, 20 parts of Radix Euonymus japonicus, 20 parts of Rosa laevigata, and 30 parts of Prunus mume.

3. A method for preparing the pharmaceutical composition for treating female infertility according to claim 1, characterized in that: The steps include: After all the drugs are mixed, they are soaked in water, decocted for the first time and filtered, the drug residues are decocted for the second time and filtered, and the filtrates are combined, concentrated, dried, and sterilized to obtain a drug composition for treating female infertility.

4. The method for preparing the pharmaceutical composition for treating female infertility according to claim 3, characterized in that: The mass ratio of the Chinese medicine to water during the soaking is 1:5-8, and the soaking time is 10-12 hours.

5. The method for preparing the pharmaceutical composition for treating female infertility according to claim 3, characterized in that: The first decoction time is 1 to 3 hours; the mass ratio of the Chinese medicine to water during the second decoction is 1:5 to 8, and the decoction time is 1 to 3 hours.

6. The method for preparing the pharmaceutical composition for treating female infertility according to claim 3, characterized in that: The pharmaceutical composition for treating female infertility can be prepared into decoction, capsule, paste, powder or pill.

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